Effects of Mechanical Stress and Phenotype Switching on Human Stem Cell-Derived Vascular Smooth Muscle Cells: Modeling Gene Regulatory Networks

机械应力和表型转换对人类干细胞衍生的血管平滑肌细胞的影响：基因调控网络建模

• 批准号: 2135907
• 负责人: Kenneth Boheler
• 金额: $ 120万
• 依托单位: Johns Hopkins University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2025-12-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2135907&HistoricalAwards=false
• 关键词: Effects; Mechanical; Stress; Phenotype; Switching

This project will reveal how human vascular smooth muscle cells (vSMCs) adapt to mechanical forces like blood pressure to switch from a state that favors the formation of new vessels to one designed to control blood flow to tissues. The mechanisms responsible for this transition in humans are poorly understood, despite the importance of this switch in human vascular development and tissue repair. Beyond scientific advances, the project will promote the training and education of undergraduate and graduate students in cell biology, physics and computational modeling that will prepare them for careers in academia and industry. Research internships with a local majority-minority magnet math-science high school will also offer training opportunities for Baltimore City students to encourage STEM-based careers. Long-term the benefits to society will likely include new approaches to model or utilize vSMCs for vessel formation and vascular repair.The goal of this research is to determine how mechanical forces regulate vSMC phenotype switching using a human vSMC surrogate model of perinatal development. The model system employed here relies on lineage-specific vSMCs differentiated from human induced pluripotent stem cells with well-defined synthetic and contractile phenotypic states. The mechanical properties and response of these vSMCs to stress/strain will be assessed using platforms that enable single cell analyses and stretching of cells in both 2D and 3D geometries. Once “tuned” to maximize internal force development, the model systems will be used with RNA-seq, bioinformatics, and computational modeling to identify key regulatory factors (e.g., transcription factors, signaling molecules) and to model regulatory networks predicted to control hPSC-vSMC responses to increased stress/strain. When coupled with ChIP-seq assays, the results will be used to decipher how mechanical forces affect transcriptional mechanisms responsible for phenotype transitions in vSMCs.This project is funded jointly by the Cellular Dynamics and Function (CDF) and Systems and Synthetic Biology (SSB) clusters of the Division of Molecular and Cellular Biosciences.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

该项目将揭示人类血管平滑肌细胞（vSMC）如何适应血压等机械力，从有利于形成新血管的状态切换到旨在控制组织血流的状态。尽管这种转变在人类血管发育和组织修复中很重要，但对人类这种转变的机制知之甚少。除了科学进步，该项目还将促进本科生和研究生在细胞生物学、物理学和计算建模方面的培训和教育，为他们在学术界和工业界的职业生涯做好准备。与当地少数民族磁铁数学科学高中的研究实习也将为巴尔的摩市的学生提供培训机会，以鼓励基于STEM的职业生涯。从长远来看，对社会的好处可能包括建立或利用vSMC进行血管形成和血管修复的新方法。本研究的目标是使用围产期发育的人类vSMC替代模型来确定机械力如何调节vSMC表型转换。本文采用的模型系统依赖于从具有明确定义的合成和收缩表型状态的人诱导多能干细胞分化的谱系特异性vSMC。这些vSMC的机械性能和对应力/应变的响应将使用能够进行单细胞分析和在2D和3D几何形状中拉伸细胞的平台进行评估。一旦“调整”以最大化内力发展，模型系统将与RNA-seq、生物信息学和计算建模一起使用，以鉴定关键调控因子（例如，转录因子、信号传导分子）和模拟预测控制hPSC-vSMC对增加的应激/应变的应答的调控网络。当与ChIP-seq测定结合时，研究结果将用于解释机械力如何影响vSMC表型转变的转录机制。该项目由细胞动力学与功能（CDF）和系统与合成生物学（SSB）联合资助该奖项反映了NSF的法定使命，并已被认为是值得通过使用评估的支持。基金会的学术价值和更广泛的影响审查标准。